Techniques in molecular biology and molecular medicine often rely on analysis of single biological molecules. Such techniques include DNA and RNA sequencing, polymorphism detection, the detection of proteins of interest, the detection of protein-nucleic acid complexes, and many others. The high sensitivity, high throughput and low reagent costs involved in single molecule analysis make it an increasingly attractive approach for a variety of detection and analysis problems in molecular medicine, from low cost genomics to high sensitivity marker analysis.
The small observation volumes used for single molecule nucleic acid sequencing and other analysis methods are typically provided by immobilizing or otherwise localizing the polymerase (or other) enzyme within an optical confinement reaction/observation region, such as an array of nanoscale wells, such as in an array of Zero Mode Waveguides (ZMWs), and delivering a template, primers, etc., to the reaction region. While these methods of nucleic acid sequencing are providing dramatic improvements in the ability to obtain sequence information, there is a need for improvements in the throughput and yield of the systems.
It is desirable to develop methods and compositions for enriching compositions for polymerase enzymes properly complexed with nucleic acid templates to provide sequencing data and to minimize the number of free polymerases in the composition that is eventually used for sequencing reactions. The present invention provides these and other features that will be apparent upon complete review of the following.